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Structure paper

TitleConnexin-46/50 in a dynamic lipid environment resolved by CryoEM at 1.9 Å.
Journal, issue, pagesNat Commun, Vol. 11, Issue 1, Page 4331, Year 2020
Publish dateAug 28, 2020
AuthorsJonathan A Flores / Bassam G Haddad / Kimberly A Dolan / Janette B Myers / Craig C Yoshioka / Jeremy Copperman / Daniel M Zuckerman / Steve L Reichow /
PubMed AbstractGap junctions establish direct pathways for cells to transfer metabolic and electrical messages. The local lipid environment is known to affect the structure, stability and intercellular channel ...Gap junctions establish direct pathways for cells to transfer metabolic and electrical messages. The local lipid environment is known to affect the structure, stability and intercellular channel activity of gap junctions; however, the molecular basis for these effects remains unknown. Here, we incorporate native connexin-46/50 (Cx46/50) intercellular channels into a dual lipid nanodisc system, mimicking a native cell-to-cell junction. Structural characterization by CryoEM reveals a lipid-induced stabilization to the channel, resulting in a 3D reconstruction at 1.9 Å resolution. Together with all-atom molecular dynamics simulations, it is shown that Cx46/50 in turn imparts long-range stabilization to the dynamic local lipid environment that is specific to the extracellular lipid leaflet. In addition, ~400 water molecules are resolved in the CryoEM map, localized throughout the intercellular permeation pathway and contributing to the channel architecture. These results illustrate how the aqueous-lipid environment is integrated with the architectural stability, structure and function of gap junction communication channels.
External linksNat Commun / PubMed:32859914 / PubMed Central
MethodsEM (single particle)
Resolution1.9 - 2.5 Å
Structure data

EMDB-22358: Connexin-46/50 in a dynamic lipid environment resolved by CryoEM at 1.9 angstroms
PDB-7jjp: Sheep Connexin-50 at 1.9 angstroms resolution by CryoEM
PDB-7jkc: Sheep Connexin-46 at 1.9 angstroms resolution by CryoEM
Method: EM (single particle) / Resolution: 1.94 Å

EMDB-22382, PDB-7jlw:
Sheep Connexin-50 at 2.5 angstroms resolution, Lipid Class 1
PDB-7jmd: Sheep Connexin-46 at 2.5 angstroms resolution, Lipid Class 1
Method: EM (single particle) / Resolution: 2.5 Å

EMDB-22390, PDB-7jm9:
Sheep Connexin-50 at 2.5 angstroms reoslution, Lipid Class 2
PDB-7jn0: Sheep Connexin-46 at 2.5 angstroms resolution, Lipid Class 2
Method: EM (single particle) / Resolution: 2.5 Å

EMDB-22391, PDB-7jmc:
Sheep Connexin-50 at 2.5 angstroms resolution, Lipid Class 3
PDB-7jn1: Sheep Connexin-46 at 2.5 angstroms resolution, Lipid Class 3
Method: EM (single particle) / Resolution: 2.5 Å

Chemicals

ChemComp-MC3:
1,2-DIMYRISTOYL-RAC-GLYCERO-3-PHOSPHOCHOLINE / phospholipid*YM

ChemComp-HOH:
WATER

Source
  • ovis aries (sheep)
  • Sheep (sheep)
KeywordsMEMBRANE PROTEIN / Connexin / Gap Junction / Nanodisc / Lipid

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